Biological effects of new chemical-mechanical caries removal products on human dental pulp stem cells.

AIM: The aim of this study was to compare the biological effects of four chemical caries removal materials and to assess their cytotoxicity using human dental pulp stem cells (hDPSCs). METHODS: The products evaluated are: 1 - Papain based product (BRIX 3000®); 2 - Papain/chloramine based products (NATURAL-CARE and Papacárie Duo®); and 3 - Chloramine based product (Cariesolut); The following in vitro experiments were carried out: IC50 measurement, cell metabolic activity (MTT) assay, cell migration, immunofluorescence experiment, cell apoptosis analysis, and reactive oxygen species (ROS) production analysis. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p<0.05). RESULTS: The IC50 values were: Brix 3000: 0.596%; Papacárie Duo: 0.052%; NATURAL CARE: 1.034%; and Cariesolut: 0.020%. The MTT assays showed non-adequate cell viability of all CMCR tested at 2% at 24, 48, and 72 h (p<0.001). The same behaviour was observed at 0.1% in the Papacárie Duo and Cariesolut groups. In contrast, 0.1% of Brix 3000 at all times and NATURAL CARE at 24h treated cells showed cell viability rates similar to the control group. At 0.01% only Brix 3000 did not show statistically significant differences at any time. Delayed cell migration was observed in all hDPSCs treated with Papacárie Duo and Cariesolut (p<0.01 and p<0.001). Phalloidin staining images showed a high confluence of cells in the presence of NATURAL CARE, similar to the control group. On the contrary, no cells were observed in Brix3000 and Cariesolut at 2% and 0.1% concentrations. Papacárie Duo showed cells at all concentrations, but hDPSCs treated at 0.01% exhibited better proliferation and spreading than those in the control group. Apoptosis assay showed that Brix 3000 at 0.1% and 0.01% had a percentage of live cells higher than 99%, with 68.4% live cells at 2%, 3.69% early apoptotic cells, and 27.9% late apoptotic cells. Conversely, the remaining materials showed abundant apoptotic cells, even at low concentrations. 0.1% and 0.01%of BRIX 3000 did not affect the ROS production levels, while 2% of BRIX 3000 counterparts significantly increased the percentage of CM-H2DCFDA positive cells. Again, all concentrations of Cariesolut showed significantly higher levels of ROS production than those observed in control cells. CONCLUSION: Our results suggest that Brix 3000 would be the most suitable material for chemical caries removal.

Biocompatibility, bioactivity and immunomodulatory properties of three calcium silicate-based sealers: an in vitro study on hPDLSCs.

OBJECTIVES: To assess the biocompatibility, bioactivity, and immunomodulatory properties of three new calcium silicate cement-based sealers: Ceraseal (CS), Totalfill BC Sealer (TFbc) and WellRoot ST (WR-ST) on human periodontal ligament stem cells (hPDLSCs). MATERIALS AND METHODS: HPDLSCs were isolated from extracted third molars from healthy patients. Eluates (1:1, 1:2, and 1:4 ratio) and sample discs of CS, TFbc and WR-ST after setting were prepared. A series of assays were performed: cell characterization, cell metabolic activity (MTT assay) cell attachment and morphology (SEM assay), cell migration (wound-healing assay), cytoskeleton organization (phaloidin-based assay); IL-6 and IL-8 release (ELISA); differentiation marker expression (RT-qPCR assay), and cell mineralization (Alizarin Red S staining). HPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media were used as a comparison. Statistical significance was established at p < 0.05. RESULTS: All the tested sealers exhibited similar results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group. CS and TFbc exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. CS and TFbc also showed a significantly higher calcified nodule formation than the negative and positive control groups. Both the marker expression and calcified nodule formation were significantly higher in CS-treated cells than TFbc treated cells. WR-ST exhibited similar results to the control group. CS and TFbc-treated cells exhibited a significant downregulation of IL-6 after 72 h of culture compared to the negative control group (p < 0.05). CONCLUSION: All the tested sealers exhibited an adequate cytocompatibility. CS significantly enhances cell differentiation by upregulating the expression of key genes associated with bone and cementum formation. Additionally, CS was observed to facilitate the mineralization of the extracellular matrix effectively. In contrast, the effects of TFbc and WR-ST on these processes were less pronounced compared to CS. Furthermore, both CS and TFbc exhibited an anti-inflammatory potential, contributing to their potential therapeutic benefits in regenerative endodontics. CLINICAL RELEVANCE: This is the first study to compare the biological properties and immunomodulatory potential of Ceraseal, Totalfill BC Sealer, and WellRoot ST. The results act as supporting evidence for their use in root canal treatment.

Integrating Endovascular Techniques into Established Open Neurosurgery Practice: A Temporal Analysis of Treatment Evolution in a Dual-Trained Neurosurgical Unit.

OBJETIVE: In Europe, units with Dual-trained Neurovascular Surgeons (DTNS) skilled in both open neurosurgery (ON) and endovascular neurosurgery (EN) are scarce. For instance, in Spain, our unit is unique within the public health system, where all neurovascular procedures are carried out by DTNS. Our study aims to evaluate the evolution in treating ruptured intracranial aneurysms (rICAs) and assess the impact of this evolution on clinical outcomes. METHODS: A retrospective cohort study was performed on rICAs treated in our unit from October 2012 to June 2023. We reviewed clinical and radiological data to analyze the evolution of ON and EN over time, as well as their impact on patient outcomes. Univariate, multivariate, and mixed-effects models were utilized to discern temporal changes. RESULTS: The modified Fisher Scale (mFS) and the modified World Federation of Neurological Surgeons scale (mWFNS) showed strong correlation with the outcome at 6 months outcomes, both with p < 0.00001. However, the surgical intervention method, ON versus EN, did not significantly affect outcomes (p > 0.85). In adjusted multivariate logistic regression, mFS (-1.579, p: 0.011) and mWFNS (-0.872, p < 0.001) maintained their significance. rICAs location was significant when comparing ON to EN p = 0.0001. A significant temporal trend favored the selection of EN p = 0.0058). Mixed-effects time series modeling indicated that while patient characteristics and rICA specifics did not predict treatment choice, the year of treatment was significantly correlated (0.161, p = 0.002). Logistic regression with interaction terms for time and treatment type did not produce significant results. CONCLUSION: Our findings suggest that despite an increased adoption of EN techniques, there has been no change in patient outcomes. Even with the rise of EN, our unit continues to perform ON for a higher proportion of rICAs than most national hospitals. We propose that a "dual approach" offers advantages in a patient individualized treatment decision protocol in the European context.

Artificial-intelligence-model to optimize biocide dosing in seawater-cooled industrial process applications considering environmental, technical, energetic, and economic aspects.

This research introduces an Artificial Intelligence (AI) based model designed to concurrently optimize energy supply management, biocide dosing, and maintenance scheduling for heat exchangers. This optimization considers energetic, technical, economic, and environmental considerations. The impact of biofilm on heat exchangers is assessed, revealing a 41% reduction in thermal efficiency and a 113% increase in flow frictional resistance of the fluid compared to the initial state. Consequently, the pump's power consumption, required to maintain hydraulic conditions, rises by 9%. The newly developed AI model detects the point at which the heat exchanger's performance begins to decline due to accumulating dirt, marking day 44 of experimentation as the threshold to commence the antifouling biocide dosing. Leveraging this AI model to monitor heat exchanger efficiency represents an innovative approach to optimizing antifouling biocide dosing and reduce the environmental impact stemming from industrial plants.

Effects of a Multicomponent Preventive Intervention in Women at Risk of Sarcopenia: A Pilot Study.

Sarcopenia is defined by the presence of decreased skeletal muscle mass, strength, and functionality in older people. Multicomponent interventions represent an alternative to non-pharmacological treatment for preventing disease progression. This study aimed to evaluate the effects of a multicomponent intervention approach in women at risk of sarcopenia. METHODS: A quasi-experimental pilot study of 12 weeks was conducted, with 24 sessions of dancing and resistance exercises and 12 sessions of nutritional education. The outcomes were changes in muscle mass, grip strength, gait speed, and body composition. The project was registered on Clinical Trials: NCT06038500 (14 September 2023). RESULTS: Twelve women aged 55-75 years participated in this study; after the intervention, changes were found in the following variables: grip strength, from 18.70 (17.98-19.23) at baseline to 21.57 (20.67-23.16) kg (p = 0.002); gait speed, from 0.95 (0.81-1.18) at baseline to 1.34 (1.20-1.47) m/s (p = 0.003); and hip circumference, from 99.75 (94.75-110.37) at baseline to 97.65 (93.92-109.50) cm (p = 0.023). Other measurements that appeared without changes were appendicular skeletal muscle mass, from 21.17 (18.58-22.33) at baseline to 20.77 (18.31-22.39) kg (p = 0.875), and the appendicular skeletal muscle mass index, from 8.64 (8.08-9.35) at baseline to 8.81 (7.91-9.38) kg/m2 (p = 0.875) after the intervention. CONCLUSIONS: The three-month multicomponent intervention in women at risk of sarcopenia improved their grip strength and gait speed.

RESPAN: an accurate, unbiased and automated pipeline for analysis of dendritic morphology and dendritic spine mapping.

Accurate and unbiased reconstructions of neuronal morphology, including quantification of dendritic spine morphology and distribution, are widely used in neuroscience but remain a major roadblock for large-scale analysis. Traditionally, spine analysis has required labor-intensive manual annotation, which is prone to human error and impractical for large 3D datasets. Previous automated tools for reconstructing neuronal morphology and quantitative dendritic spine analysis face challenges in generating accurate results and, following close inspection, often require extensive manual correction. While recent tools leveraging deep learning approaches have substantially increased accuracy, they lack functionality and useful outputs, necessitating additional tools to perform a complete analysis and limiting their utility. In this paper, we describe Restoration Enhanced SPine And Neuron (RESPAN) analysis, a new comprehensive pipeline developed as an open-source, easily deployable solution that harnesses recent advances in deep learning and GPU processing. Our approach demonstrates high accuracy and robustness, validated extensively across a range of imaging modalities for automated dendrite and spine mapping. It also offers extensive visual and tabulated data outputs, including detailed morphological and spatial metrics, dendritic spine classification, and 3D renderings. Additionally, RESPAN includes tools for validating results, ensuring scientific rigor and reproducibility.

Cornerball: a new alternative sport proposal for school physical education.

Physical Education has had to evolve and change throughout history to adapt to the demands of society. As a result, teachers have had to seek pedagogical alternatives to ensure that students are active, motivated, and engaged in the classroom. This approach allows for the development of motor, cognitive, and socio-emotional skills in students, ultimately contributing to the holistic development of the individual, which is the primary goal of education. Out of this intrinsic need for Physical Education, Alternative Sports have emerged, providing opportunities for different types of learning compared to more traditional sports. Cornerball, a hybrid between split-court and wall-based sports, played in a somewhat unique playing area-a 90° corner, is introduced with the aim of offering a new Alternative Sport. The objective of this descriptive study is to present a new pedagogical proposal designed for the educational context, highlighting its characteristics and fundamental aspects to consider, such as rules, the playing field, participants, and methodological strategies for its implementation within a Physical Education classroom. Therefore, the aims and purposes of this work are to describe a new sports game so that in the future, more detailed and specific empirical studies can be proposed.

Test-retest reliability of putting-related variables in medium-to-high handicap golf players.

This manuscript aims to study the reliability of different variables related to performance and acceleration during the golf putt in players with medium-to-high handicaps and to determine the number of attempts necessary to find reliable values for these variables. Eight males and two females [55.67 (13.64) years, 78.4 (11.4) kg, 1.75 (7.95) m] participated in two experimental sessions separated by one week. In these sessions, they performed three blocks of 10 putts trying to stop the golf ball at the center of a dartboard painted 2 m away. The performance was assessed depending on the area of the dartboard where the ball stopped, and the acceleration signals were acquired using the Xsens Dot. The results showed that to evaluate performance, 18 trials were necessary to reach reliable values using the 0-10 scoring system, and 28 trials were necessary for the 0-3 scoring system. Regarding the reliability of the accelerometer-related variables, 7 attempts were necessary to obtain good-to-excellent reliability values for most of the variables. It could be concluded that putting in medium-to-high handicap golf players can be reliably measured using the abovementioned protocol.

In Silico Chemical Experiments in the Age of AI: From Quantum Chemistry to Machine Learning and Back.

Computational chemistry is an indispensable tool for understanding molecules and predicting chemical properties. However, traditional computational methods face significant challenges due to the difficulty of solving the Schrödinger equations and the increasing computational cost with the size of the molecular system. In response, there has been a surge of interest in leveraging artificial intelligence (AI) and machine learning (ML) techniques to in silico experiments. Integrating AI and ML into computational chemistry increases the scalability and speed of the exploration of chemical space. However, challenges remain, particularly regarding the reproducibility and transferability of ML models. This review highlights the evolution of ML in learning from, complementing, or replacing traditional computational chemistry for energy and property predictions. Starting from models trained entirely on numerical data, a journey set forth toward the ideal model incorporating or learning the physical laws of quantum mechanics. This paper also reviews existing computational methods and ML models and their intertwining, outlines a roadmap for future research, and identifies areas for improvement and innovation. Ultimately, the goal is to develop AI architectures capable of predicting accurate and transferable solutions to the Schrödinger equation, thereby revolutionizing in silico experiments within chemistry and materials science.

Assessment of the anti-inflammatory and biological properties of Bioroot Flow: A novel bioceramic sealer.

INTRODUCTION: BioRoot Flow (BRF) is a novel premixed bioceramic sealer indicated for endodontic treatments, but the biological and immunomodulatory effects of this endodontic sealer on human periodontal ligament stem cells (hPDLSCs) have not been elucidated. METHODS: To ascertain the biological impact of BRF, TotalFill BC Sealer (TFbc), and AH Plus (AHP) on human Periodontal Ligament Stem Cells (hPDLSCs), assessments were conducted to evaluate the cytocompatibility, cellular proliferation, migratory capacity, osteo/cementogenic differentiation potential, the ability to form mineralized nodules, and the immunomodulatory characteristics of hPDLSCs following treatment with these endodontic sealers. RESULTS: Biological assays showed adequate cell metabolic activity and cell migration in BRF, while SEM assay evidenced that TFbc and BRF groups demonstrated a superior cell adhesion process, including substrate adhesion, cytoskeleton development, and spreading on the niche-like structures of the cement as compared to the AHP group. TFbc and BRF-treated groups exhibited a significantly lower IL6 and IL8 production than AHP (* p <.05). The bioceramic sealers stimulated heightened expression of BSP, CEMP-1, and CAP genes within a 7-14 day period. Notably, BRF and TFbc demonstrated a significant enhancement in the mineralization of hPDLSCs when compared to the negative control. Among these, cells treated with BRF showed a more substantial accumulation of calcium (*** p < .001). CONCLUSIONS: Taken together, these findings indicate that BRF can potentially enhance cell differentiation by promoting the expression of essential genes related to bone and cement formation. In addition, BRF and TFbc displayed anti-inflammatory effects.

Comparative Cytotoxicity of Menthol and Eucalyptol: An In Vitro Study on Human Gingival Fibroblasts.

The aim of this study was to assess the influence of eucalyptol and menthol on the cell viability, migration, and reactive oxygen species production of human gingival fibroblasts (GFs) in vitro. Three different concentrations of eucalyptol and menthol were prepared following ISO 10993-5 guidelines (1, 5, and 10 mM). GFs were isolated from extracted teeth from healthy donors. The following parameters were assessed: cell viability via MTT, Annexin-V-FITC and 7-AAD staining, and IC50 assays; cell migration via horizontal scratch wound assay; and cell oxidative stress via reactive oxygen species assay. Data were analyzed using one-way ANOVA and Tukey's post hoc test. Statistical significance was established at p < 0.05. Eucalyptol and Menthol exhibited high cytotoxicity on gingival fibroblasts, as evidenced by cytotoxicity assays. Eucalyptol showed lower levels of cytotoxicity than menthol, compared to the control group. The cytotoxicity of the tested substances increased in a concentration-dependent manner. The same occurred in a time-dependent manner, although even 10 min of exposure to the tested substances showed a high cytotoxicity to the GFs. Commercially available products for oral application with these substances in their composition should be tested for cytotoxicity before their use.

[Economic burden assessment for the management of asthma patients at Mexico's National Institute for Respiratory Diseases]. / Evaluación de la carga económica para el tratamiento de pacientes con asma en el Instituto Nacional de Enfermedades Respiratorias de México.

OBJETIVO: Determinar la carga económica anual del asma, desde una perspectiva institucional y con base en la clasificación recomendada por GINA, en una cohorte retrospectiva de adultos atendidos en el Instituto Nacional de Enfermedades Respiratorias (INER) de México. MÉTODOS: Estudio observacional, longitudinal y retrospectivo, llevado a cabo a partir de la información recabada de 247 pacientes femeninas con asma. Se estimaron los costos directos anuales: visitas, pruebas de laboratorio, tratamiento farmacológico y de las crisis o exacerbaciones, para determinar la carga anual de la enfermedad desde una perspectiva institucional, y según la clasificación de la Iniciativa Global para el Asma. RESULTADOS: El costo promedio anual fue de $43,813,92, que aumentó en relación con la necesidad de aumento de dosis de corticoides inhalados y beta-agonistas de acción prolongada. El costo promedio de la consulta médica fue de $2004.57, $982.82 por gestión de crisis y $2645.95 por pruebas de laboratorio. El tratamiento farmacológico representó la principal carga económica, con un costo promedio anual de $38,180.58. CONCLUSIONES: Los resultados resaltan una carga económica del asma estimada en un costo anual por paciente de $43,813.92 MXN (DE=93,348.85), en el contexto del tercer nivel de atención en el sistema de salud público mexicano. La gravedad del asma, los tratamientos y los biológicos fueron los principales factores que aumentaron los costos directos de la atención.

OBJECTIVE: Determine the annual economic burden of the disease from an institutional perspective and based on GINA's recommended classification in a retrospective cohort of adults treated at Instituto Nacional de Enfermedades Respiratorias (INER) of Mexico City. METHODS: A retrospective, longitudinal observational study comprised by data from 247 female asthma patients, annual direct costs were estimated including: visits, laboratory tests, pharmacological treatment and management of crisis or exacerbations, to determine the annual burden of the disease from an institutional perspective and according to Global Initiative for Asthma classification. RESULTS: The average annual cost was $43,813.92, which increased in relation to the need of inhaled corticosteroids and long-acting beta agonists dosage increase. The average doctor's appointment cost was $2,004.57, $982.82 for crisis management and $2,645.95 for laboratory testing. Pharmacological treatment represented the main economic burden with an annual average cost of $38,180.58. CONCLUSIONS: The results highlight an economic burden of asthma estimated at an annual cost per patient of $43,813.92 MXN (SD=93,348.85) in the context of the third level of care in the Mexican public health system. The asthma severity and treatments such as biologics were the main factors that increased direct costs of care.

Evaluation of particulate organic carbon from MODIS-Aqua in a marine-coastal water body.

La Paz Bay (LPB) in Mexico is one of the largest marine-coastal bodies of water in the Gulf of California (GC) and is ecologically important for the feeding, reproduction, and refuge of marine species. Although particulate organic carbon (POC) is an important reservoir of oceanic carbon and an indicator of productivity in the euphotic zone, studies in this region are scarce. This study evaluates the performance of satellite-derived POC in LPB from January 2003 to December 2020. The metrics obtained for COP ( RMSE = 33.8 mg m - 3 ; P bias = 29.6 % y r P = 0.4 con p < 0.05 ), Chla-a ( RMSE = 0.23 mg m - 3 ; P bias = - 4.3 % y r P = 0.94 con p < 0.05 ), and SST ( RMSE = 2 . 3 ∘ C ; P bias = - 2.2 % y r P = 0.92 con p < 0.05 ) establish that although in some cases there was a slight over/underestimation, the satellite estimates consistently represent the variability and average values measured in situ. On the other hand, the spatio-temporal analysis of the POC allowed us to identify two seasons with their respective transition periods and five subregions in which the POC is characterized by having its maximum variability; two of these coincide with the locations of the eddies reported for the winter and summer seasons in the LPB, while the following three are located: one in the coastal zone and in the two areas in which the LPB interacts with the GC. The associations, variability nodes, and multiple linear regression analysis suggest that POC fluctuations in the LPB respond mainly to biological processes and, to some extent, to the seasonality of SST and wind. Finally, our results justify the use of the MODIS-Aqua satellite POC for studies in marine-coastal water bodies with similar characteristics to the LPB and suggest that this water body can be considered a reservoir for the marine region of northwestern Mexico.

A comparative study of CNN-capsule-net, CNN-transformer encoder, and Traditional machine learning algorithms to classify epileptic seizure.

INTRODUCTION: Epilepsy is a disease characterized by an excessive discharge in neurons generally provoked without any external stimulus, known as convulsions. About 2 million people are diagnosed each year in the world. This process is carried out by a neurological doctor using an electroencephalogram (EEG), which is lengthy. METHOD: To optimize these processes and make them more efficient, we have resorted to innovative artificial intelligence methods essential in classifying EEG signals. For this, comparing traditional models, such as machine learning or deep learning, with cutting-edge models, in this case, using Capsule-Net architectures and Transformer Encoder, has a crucial role in finding the most accurate model and helping the doctor to have a faster diagnosis. RESULT: In this paper, a comparison was made between different models for binary and multiclass classification of the epileptic seizure detection database, achieving a binary accuracy of 99.92% with the Capsule-Net model and a multiclass accuracy with the Transformer Encoder model of 87.30%. CONCLUSION: Artificial intelligence is essential in diagnosing pathology. The comparison between models is helpful as it helps to discard those that are not efficient. State-of-the-art models overshadow conventional models, but data processing also plays an essential role in evaluating the higher accuracy of the models.

Dietary Concentrate Supplementation Increases Milk Production and Reduces Predicted Greenhouse Gas Emissions Intensity in Pasture-based Commercial Dairy Farms.

Controlled studies have extensively documented that concentrate supplements typically increase enteric methane (CH4) emissions and milk yield and reduce emissions per unit of milk produced and dry matter intake. However, there have been no studies conducted to determine the effect of concentrate on predicted greenhouse gas emissions from dairy farms representing the Australian pasture-based farming system. Thus, this study sought to determine how dietary concentrate supplementation affects enteric and manure CH4, and N2O of Australian pasture-based dairy farms. The Australian Dairy Carbon Calculator was used, which incorporates emission factors and methodologies used in the National Greenhouse Gas Inventory as reported to the International Panel on Climate Change. Primary data were collected and analyzed from 120 commercial farms in Australia's major dairy regions. Then the farms were divided into 4 groups based on their dietary concentrate supplementation: ≤ 1 (low; 15 farms), 1-2 (moderate;35 farms), 2-3 (high; 35 farms), and ≥ 3 (very high; 35 farms) ton (t) of concentrate dry matter per cow per year. Sources of greenhouse gas emissions were CO2 from concentrate production, enteric CH4, and manure CH4 and N2O. Total dry matter intake, milk yield, and daily enteric CH4 production (g/day) quadratically increased with concentrate level, whereas greenhouse gas emission intensity of milk production (kg CO2eq/kg fat and protein corrected milk) decreased by 14% for farms supplementing with ≥ 3 tDM/cow per year compared with those supplementing with ≤ 1 tDM/cow per year of dietary concentrate. The emissions of N2O and CH4 from manure increased quadratically and linearly, respectively, with the increasing supplementation of concentrate. Farms supplementing 2-3 tDM/cow per year showed substantial increases in gross income, gross margin, earnings before interest and tax, and net income ($/cow/year) compared with those supplementing of ≤ 1, 1-2, and ≥ 3 tDM/cow per year. In conclusion, increasing dietary concentrate supplementation for dairy cows resulted in increased milk production per cow, reduced greenhouse gas emissions per unit of milk produced, and increased income and profit. However, a comprehensive life cycle assessment study is needed to account for carbon sequestration by other farm components such as pastures and trees, which were not considered in the present study. In addition, the present study was based on modeling and did not gather ground truth information for DMI, digestibility, crude protein, and urinary and fecal N excretion. Therefore, data should be interpreted with caution, and studies gathering such information are encouraged.

Premixed calcium silicate-based ceramic sealers promote osteogenic/cementogenic differentiation of human periodontal ligament stem cells: A microscopy study.

To evaluate the effects of premixed calcium silicate based ceramic sealers on the viability and osteogenic/cementogenic differentiation of human periodontal ligament stem cells (hPDLSCs). The materials evaluated were TotalFill BC Sealer (TFbc), AH Plus Bioceramic Sealer (AHPbc), and Neosealer Flo (Neo). Standardized discs and 1:1, 1:2, and 1:4 eluates of the tested materials were prepared. The following in vitro experiments were carried out: ion release, cell metabolic activity 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, cell migration, immunofluorescence experiment, cell attachment, gene expression, and mineralization assay. Statistical analyses were performed using one-way ANOVA followed by Tukey's post hoc test (p < .05). Increased Ca2+ release was detected in TFbc compared to AHPbc and Neo (*p < .05). Biological assays showed a discrete cell metabolic activity and cell migration in Neo-treated cell, whereas scanning electronic microscopy assay exhibited that TFbc group had a better cell adhesion process of substrate attachment, spreading, and cytoskeleton development on the niche-like structures of the cement than AHPbc and Neo. The sealers tested were able to induce overexpression of the CEMP-1, ALP, and COL1A1 genes in the first days of exposure, particularly in the case of TFbc (***p < .001). All materials tested significantly increased the mineralization of hPDLSCs when compared to the negative control, although more pronounced calcium deposition was observed in the TFbc-treated cells (***p < .001). Our results suggested that TFbc promotes cell differentiation, both by increasing the expression of key osteo/odontogenic genes and by promoting mineralization of the extracellular matrix, whereas this phenomenon was less evident in Neo and AHPbc. RESEARCH HIGHLIGHTS: TFbc group had a better cell adhesion process of substrate attachment, spreading, and cytoskeleton development on the niche-like structures of the cement than AHPbc and Neo. The sealers tested were able to induce overexpression of the CEMP-1, ALP, and COL1A1 genes in the first days of exposure, particularly in the case of TFbc. All materials tested significantly increased the mineralization of hPDLSCs when compared to the negative control, although more pronounced calcium deposition was observed in the TFbc-treated cells.

Comparative bioactivity and immunomodulatory potential of the new Bioroot Flow and AH Plus Bioceramic sealer: An in vitro study on hPDLSCs.

OBJECTIVES: To evaluate the cytocompatibility, bioactivity, and anti-inflammatory potential of the new pre-mixed calcium silicate cement-based sealers Bioroot Flow (BrF) and AH Plus Bioceramic Sealer (AHPbcs) on human periodontal ligament stem cells (hPDLSCs) compared to the epoxy resin-based sealer AH Plus (AHP). MATERIALS AND METHODS: Standardized discs and 1:1, 1:2, and 1:4 eluates of BrF, AHPbcs and AHP after setting were prepared. The following assays were performed: cell attachment and morphology via SEM, cell viability via a MTT assay, cell migration/proliferation via a wound-healing assay, cytoskeleton organization via immunofluorescence staining; cytokine release via ELISA; osteo/cemento/odontogenic marker expression via RT-qPCR, and cell mineralized nodule formation via Alizarin Red S staining. HPDLSCs were isolated from extracted third molars from healthy patients. Comparisons were made with hPDLSCs cultured in unconditioned (negative control) or osteogenic (positive control) culture media. Statistical significance was established at p < 0.05. RESULTS: Both BrF and AHPbcs showed significantly positive results in the cytocompatibility assays (cell metabolic activity, migration, attachment, morphology, and cytoskeleton organization) compared with a negative control group, while AHP showed significant negative results. BrF exhibited an upregulation of at least one osteo/cementogenic marker compared to the negative and positive control groups. BrF showed a significantly higher calcified nodule formation than AHPbcs, the negative and positive control groups, while AHPbcs was higher than the negative control group. Both were also significantly higher than AHP group. CONCLUSION: BrF and AHPbcs exhibit adequate and comparable cytocompatibility on hPDLSCs. BrF also promoted the osteo/cementogenic differentiation of hPDLSCs. Both calcium silicate-based sealers favored the downregulation of the inflammatory cytokine IL-6 and the calcified nodule formation from hPDLSCs. BrF exerted a significantly higher influence on cell mineralization than AHPbcs. CLINICAL RELEVANCE: This is the first study to elucidate the biological properties and immunomodulatory potential of Bioroot Flow and AH Plus Bioceramic Sealer. The results act as supporting evidence for their use in root canal treatment.